Cholesterol FS*

Diagnostic reagent for quantitative in vitro determination of cholesterol in serum or plasma on photometric
systems

Order Information Storage Instructions and Reagent Stability

Cat. No. Kit size Reagent and standard are stable up to the end of the indicated
1 1350 99 10 021 R 5x 25 mL + 1 x 3 mL Standard month of expiry, if stored at 2 – 8°C, protected from light and
1 1350 99 10 026 R 6x 100 mL contamination is avoided. Do not freeze the reagents!
1 1350 99 10 023 R 1x 1000 mL
1 1300 99 10 030 6x 3 mL Standard Note: It has to be mentioned, that the measurement is not
influenced by occasionally occurring color changes, as long as the
Summary [1,2] absorbance of the reagent is < 0.3 at 546 nm.
Cholesterol is a component of cell membranes and a precursor for Warnings and Precautions
steroid hormones and bile acids synthesized by body cells and 1. The reagent contains sodium azide (0.95 g/L) as preservative.
absorbed with food. Cholesterol is transported in plasma via Do not swallow! Avoid contact with skin and mucous
lipoproteins, namely complexes between lipids and apolipoproteins. membranes.
There are four classes of lipoproteins: high density lipoproteins 2. Standard: Warning. H317 May cause an allergic skin reaction.
(HDL), low density lipoproteins (LDL), very low density lipoproteins H319 Causes serious eye irritation. P264 Wash hands and
(VLDL) and chylomicrons. While LDL is involved in the cholesterol face thoroughly after handling. P280 Wear protective
transport to the peripheral cells, HDL is responsible for the gloves/protective clothing/eye protection/face protection.
cholesterol uptake from the cells. The four different lipoprotein P302+P352 If on skin: Wash with plenty of soap and water.
classes show distinct relationship to coronary atherosclerosis. LDL- P337+P313 If eye irritation persists: Get medical
cholesterol (LDL-C) contributes to atherosclerotic plaque formation advice/attention.
within the arterial intima and is strongly associated with coronary 3. In very rare cases, samples of patients with gammopathy
heart disease (CHD) and related mortality. Even with total might give falsified results [8].
cholesterol within the normal range an increased concentration of 4. N-acetylcysteine (NAC), acetaminophen and metamizole
LDL-C indicates high risk. HDL-C has a protective effect impeding medication leads to falsely low results in patient samples.
plaque formation and shows an inverse relationship to CHD 5. Please refer to the safety data sheets and take the necessary
prevalence. In fact, low HDL-C values constitute an independent precautions for the use of laboratory reagents. For diagnostic
risk factor. The determination of the individual total cholesterol (TC) purposes, the results should always be assessed with the
level is used for screening purposes while for a better risk patient’s medical history, clinical examinations and other
assessment it is necessary to measure additionally HDL-C and findings.
LDL-C. 6. For professional use only!
In the last few years several controlled clinical trials using diet, life
style changes and/or different drugs (especially HMG CoA Waste Management
reductase inhibitors [statins]) have demonstrated that lowering total Please refer to local legal requirements.
cholesterol and LDL-C levels reduce drastically CHD risk [2]. Reagent Preparation
Method The reagent and the standard are ready to use.
“CHOD-PAP”: enzymatic photometric test Materials required but not provided
NaCl solution 9 g/L
Principle General laboratory equipment
Determination of cholesterol after enzymatic hydrolysis and
oxidation [3,4]. The colorimetric indicator is quinoneimine which is Specimen
generated from 4-aminoantipyrine and phenol by hydrogen Serum, heparin plasma or EDTA plasma
peroxide under the catalytic action of peroxidase (Trinder’s Stability [6]: 7 days at 20 – 25°C
reaction) [3]. 7 days at 4 – 8°C
CHE 3 months at –20°C
Cholesterol ester + H2 O Cholesterol + Fatty acid
Discard contaminated specimens! Freeze only once!
Cholesterol + O2 CHO Cholesterol-3-one + H2O2
Assay Procedure
2 H2O2 + 4-Aminoantipyrine + Phenol POD Quinoneimine + 4 H2O Application sheets for automated systems are available on
request.
Reagents Wavelength 500 nm, Hg 546 nm
Optical path 1 cm
Components and Concentrations Temperature 20 – 25°C/37°C
Reagent: Measurement Against reagent blank
Good's buffer pH 6.7 50 mmol/L
Phenol 5 mmol/L Blank Sample or standard
4-Aminoantipyrine 0.3 mmol/L Sample or standard - 10 µL
Cholesterol esterase (CHE)  200 U/L Dist. water 10 µL -
Cholesterol oxidase (CHO)  100 U/L Reagent 1000 µL 1000 µL
Peroxidase (POD)  3 kU/L Mix, incubate for 10 min. at 20 – 25°C or for 5 min. at 37°C. Read
Standard: 200 mg/dL (5.2 mmol/L) absorbance within 60 min against reagent blank.

Cholesterol FS – Page 1 * fluid stable

Calculation Reference Range [5]
With standard or calibrator Desirable  200 mg/dL (5.2 mmol/L)
Borderline high risk 200 – 240 mg/dL (5.2 – 6.2 mmol/L)
A Sample
Cholesterol [mg / dL]  x Conc. Std / Cal [mg / dL] High risk > 240 mg/dL (> 6.2 mmol/L)
A Std / Cal
Each laboratory should check if the reference ranges are
Conversion factor transferable to its own patient population and determine own
Cholesterol [mg/dL] x 0.02586 = Cholesterol [mmol/L] reference ranges if necessary.

Calibrators and Controls Clinical Interpretation

For the calibration of automated photometric systems, DiaSys The European Task Force on Coronary Prevention recommends to
TruCal U calibrator is recommended. The assigned values of the lower TC concentration to less than 190 mg/dL (5.0 mmol/L) and
calibrator have been made traceable to the reference method gas LDL-cholesterol to less than 115 mg/dL (3.0 mmol/L) [2].
chromatography-isotope dilution mass spectrometry (GC-IDMS).
DiaSys TruLab N and P or TruLab L controls should be assayed for Literature
internal quality control. Each laboratory should establish corrective 1. Rifai N, Bachorik PS, Albers JJ. Lipids, lipoproteins and
action in case of deviations in control recovery. apolipoproteins. In: Burtis CA, Ashwood ER, editors. Tietz
rd
Cat. No. Kit size Textbook of Clinical Chemistry. 3 ed. Philadelphia: W.B
TruCal U 5 9100 99 10 063 20 x 3 mL Saunders Company; 1999. p. 809-61.
5 9100 99 10 064 6 x 3 mL 2. Recommendation of the Second Joint Task Force of European
TruLab N 5 9000 99 10 062 20 x 5 mL and other Societies on Coronary Prevention. Prevention of
5 9000 99 10 061 6 x 5 mL coronary heart disease in clinical practice. Eur Heart J
TruLab P 5 9050 99 10 062 20 x 5 mL 1998;19: 1434-503.
5 9050 99 10 061 6 x 5 mL
3. Artiss JD, Zak B. Measurement of cholesterol concentration.
TruLab L Level 1 5 9020 99 10 065 3 x 3 mL
TruLab L Level 2 5 9030 99 10 065 3 x 3 mL In: Rifai N, Warnick GR, Dominiczak MH, eds. Handbook of
lipoprotein testing. Washington: AACC Press, 1997: 99–114.
4. Deeg R, Ziegenhorn J. Kinetic enzymatic method for
Performance Characteristics automated determination of total cholesterol in serum. Clin
Measuring range Chem 1983;29:1798-802.
5. Schaefer EJ, McNamara J. Overview of the diagnosis and
The test has been developed to determine cholesterol
treatment of lipid disorders. In: Rifai N, Warnick GR,
concentrations within a measuring range from 3 - 750 mg/dL
Dominiczak MH, eds. Handbook of lipoprotein testing.
(0.08 - 19.4 mmol/L). When values exceed this range samples
Washington: AACC press, 1997:25–48.
should be diluted 1 + 4 with NaCl solution (9 g/L) and the result
6. Guder WG, Zawta B et al. The Quality of Diagnostic Samples.
multiplied by 5. st
1 ed. Darmstadt: GIT Verlag; 2001. p. 22-3.
Specificity/Interferences 7. Young DS. Effects of Drugs on Clinical Laboratory Tests. 5th
No interference was observed by ascorbic acid up to 5 mg/dL, ed. Volume 1 and 2. Washington, DC: The American
bilirubin up to 20 mg/dL, hemoglobin up to 200 mg/dL and lipemia Association for Clinical Chemistry Press 2000.
up to 2000 mg/dL triglycerides. For further information on interfering 8. Bakker AJ, Mücke M. Gammopathy interference in clinical
substances refer to Young DS [7]. chemistry assays: mechanisms, detection and prevention.
ClinChemLabMed 2007;45(9):1240–1243.
Sensitivity/Limit of Detection
The lower limit of detection is 3 mg/dL (0.08 mmol/L). Manufacturer
Precision (at 37°C) DiaSys Diagnostic Systems GmbH
IVD
Alte Strasse 9 65558 Holzheim Germany
Intra-assay precision Mean SD CV
n = 20 [mg/dL] [mg/dL] [%]
Sample 1 108 1.76 1.62
Sample 2 236 1.45 0.61
Sample 3 254 1.57 0.62

Inter-assay precision Mean SD CV

n = 20 [mg/dL] [mg/dL] [%]
Sample 1 104 1.19 1.14
Sample 2 211 2.57 1.22
Sample 3 245 2.28 0.93
Method Comparison
A comparison of DiaSys Cholesterol FS (y) with a commercially
available test (x) using 78 samples gave following results:
y = 1.00 x – 2.50 mg/dL; r = 0.995

Cholesterol FS – Page 2 844 1350 10 02 00 July 2016/11